卡斯帕酶-3抑制剂的高通量筛选及活性化合物F03ZA-575的初步研究

目的筛选并初步研究微生物来源的卡斯帕酶-3(Caspases-3)抑制剂活性化合物。方法使用E.coli异源表达的卡斯帕酶-3作为靶酶,建立基于Caspase-3酶抑制剂的体外高通量筛选模型并对微生物来源的共计10026个次级代谢产物提取物进行了筛选。使用有机溶剂萃取、硅胶柱和LH-20柱层析、高压液相分离等方法从代谢产物中分离活性化合物并经各种理化性质及NMR分析等对活性化合物的结构进行确定。结果从10026个微生物来源的代谢产物中筛选获得了10个阳性样品。其中,从1株真菌的代谢产物中分离得到的化合物F03ZA-575对Caspase-3酶显示出较强的抑制活性,结构解析确认该化合物与Duclauxin同质。结论该化合物对Caspase-3酶的抑制活性将有助于揭示其抗肿瘤作用的机制。

High throughput screening of caspase-3 inhibitors and the primary study of active compound F03ZA-575

Objective Screening of new Caspase-3 inhibitors from the metabolites of microorganism for the discovery of new active compounds.Methods A high throughput inhibitors screening method had been set up with the E.coli heterologous expressed human Caspase-3 as target and 10026 extracts of microbial secondary metabolite had been screened. The active compound was isolated from the metabolites by solvent extraction, silica column and LH-20 chromatography, HPLC purification etc. The structure of active compound was elucidated by means of physico-chemical properties and NMR analysis.Results Ten positive samples had been picked up from 10026 extracts of microbial secondary metabolite. One active compound named F03ZA-575 was isolated from the metabolites of one strain of fungi showed strong inhibitory activity to Caspase-3. The structure of this compound was elucidated to be identical with Duclauxin.Conclusion Its inhibitory activity to Caspase-3 was first reported and it may be helpful to clarify its anti-tumor mechanism.